Minami Nihon Rakuno Kyodo Co.

Miyakonojō, Japan

Minami Nihon Rakuno Kyodo Co.

Miyakonojō, Japan
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Biswas G.,University of Miyazaki | Korenaga H.,University of Miyazaki | Nagamine R.,University of Miyazaki | Takayama H.,University of Miyazaki | And 6 more authors.
Fish and Shellfish Immunology | Year: 2013

Cytokine responses in the Japanese pufferfish (Takifugu rubripes) head kidney (HK) cells to heat-killed lactic acid bacteria probiotics isolated from the Mongolian dairy products were investigated by transcriptomic examination. The HK cells were incubated with two heat-killed bacteria, namely Lactobacillus paracasei spp. paracasei (strain 06TCa22) and L. plantarum (strain 06CC2) and the responses of 16 cytokine genes at 0 (control), 1, 4, 8, 12, 24 and 48. h post-stimulation were assayed by multiplex RT-PCR analysis (GenomeLab Genetic Analysis System, GeXPS; Beckman Coulter, Inc.). The 16 genes included in the assay were pro-inflammatory cytokines (IL-1β, IL-6, IL-17A/F-3, TNF-α and TNF-N), cell-mediated immune regulators (IL-12p35, IL-12p40 and IL-18), antiviral (I-IFN-1 and IFN-γ) and other regulatory (IL-2, IL-7, IL-15, IL-21, IL-10 and TGF-β1) cytokines. Despite the differences in the transcriptional profiles, expression of all the cytokines tested here was significantly elevated by both the probiotic bacterial stimulants compared with the unstimulated control. Therefore, this in vitro study has demonstrated the modulation of cytokine defense mechanisms in the HK cells by the two heat-killed probiotics indicating their potentiality as novel immunostimulants to fish. However, strain-dependent varied expression of important cytokines (cell-mediated immune regulators, antiviral and anti-inflammatory cytokines) suggests better efficacy of L. paracasei spp. paracasei strain as fish immunostimulant. Further in vivo studies to elucidate the cytokine regulation networks will validate our present observations. A careful evaluation of ant-inflammatory properties may be undertaken using single strain to affirm the immunostimulatory capability. Moreover, application timings and frequency to assess the longevity of immunostimulant effects and to make the application cost-effective need to be evaluated before any practical use in aquaculture. © 2013 Elsevier Ltd.


Biswas G.,University of Miyazaki | Korenaga H.,University of Miyazaki | Nagamine R.,University of Miyazaki | Kawahara S.,University of Miyazaki | And 6 more authors.
Fish and Shellfish Immunology | Year: 2013

With the aim of evaluating the effect of a Mongolian dairy product derived Lactobacillus paracasei spp. paracasei (strain 06TCa22) (Lpp) on the cytokine-mediated immune responses to Vibrio harveyi infection, we examined 16 cytokine expressions in the Japanese pufferfish, Takifugu rubripes. Fish were orally treated with the heat-killed Lpp at 1mgg-1bodyweightd-1 for 3 days. At 24h posttreatment, fish were infected by an intramuscular injection of 0.1mL V.harveyi bacterial suspension (108cfumL-1). Additionally, superoxide anion production (SAP) and phagocytic activity (PA) of head kidney cells were assessed during 120h postinfection period. Significant up-regulation of pro-inflammatory (IL-1β, IL-6, IL-17A/F-3, TNF-α and TNF-N), cell-mediated immune inducing (IL-12p35, IL-12p40 and IL-18), antiviral/intra-cellular pathogen killing (I-IFN-1 and IFN-γ), anti-inflammatory (IL-10) and lymphocyte agonistic (IL-2, IL-7, IL-15, IL-21 and TGF-β1) cytokines was observed in the treated fish compared to control ones during the pathogen infection. Furthermore, significantly increased SAP and PA (P<0.01; 0.05) were recorded in the treated fish compared to untreated fish. These results suggest the beneficial role of Lpp in enhancement of cytokine-mediated immunity in the Japanese pufferfish against V.harveyi infection and application of this product as a potential fish immunostimulant. © 2013 Elsevier Ltd.


Matsusaki T.,Minami Nihon Rakuno Kyodo Co. | Takeda S.,Minami Nihon Rakuno Kyodo Co. | Takeda S.,Azabu University | Takeshita M.,Minami Nihon Rakuno Kyodo Co. | And 7 more authors.
International Immunopharmacology | Year: 2016

We previously found that Lactobacillus plantarum strain 06CC2 showed probiotic potential, and its oral administration effectively induced Th1 cytokine production and activated the Th1 immune response associated with intestinal immunity in mice. In this study, to evaluate its potential as a versatile oral adjuvant for treatment of viral infection, we assessed the immunomodulatory activity of 06CC2 on murine cutaneous herpes simplex virus type 1 (HSV-1) infection, in which a major immune defense system is a delayed-type hypersensitivity (DTH) reaction based on activation of the Th1 immune response, in relation to its oral efficacy for alleviation of herpetic symptoms. In the HSV-1 infection model, oral administration of 06CC2 (20 mg/mouse) twice daily for seven days starting two days before infection was significantly effective in delaying the development of skin lesions in the early phase of infection and reducing virus yields in the brain on day 4 after infection. In addition, 06CC2 significantly augmented the DTH reaction to inactivated HSV-1 antigen and elevated interferon (IFN)-γ production by HSV-1 antigen from splenocytes. On day 2, natural killer (NK) cell activity was significantly elevated, and the elevation was still observed on day 4. Furthermore, gene expressions of interleukin-12 receptor β2 and IFN-γ in Peyer's patches were augmented on day 4 by 06CC2 administration. Thus, 06CC2 was suggested to alleviate herpetic symptoms in mice in correlation with augmentation of the Th1 immune responses associated with NK cell activity through intestinal immunity. Strain 06CC2 may be a versatile oral adjuvant to activate Th1 immune response. © 2016 Elsevier B.V.


Takeda S.,Minami Nihon Rakuno Kyodo Co. | Fujimoto R.,Minami Nihon Rakuno Kyodo Co. | Takenoyama S.-I.,Minami Kyushu University | Takeshita M.,Minami Nihon Rakuno Kyodo Co. | And 5 more authors.
Food Science and Technology Research | Year: 2013

We screened suitable lactic acid bacteria (LAB) strains for use in fermented dairy products by analyzing milk fermented with 10 probiotic LAB strains from Mongolian dairy products. Lactobacillus paracasei paracasei strain 06TCa19 was selected because of its favorable effects on pH, lactic acid production, and viable bacterial numbers after fermentation of skim milk. Then, we prepared 06TCa19 and control fermented milks and conducted a randomized, double-blind crossover study with 46 healthy women to determine the effects of the strain on human defecation. The ingestion of 06TCa19 fermented milk improved the subjects' fecal characteristics, including shape and color. Analysis of stool samples from 8 subjects revealed that l-lactic acid levels and Lactobacillus and Bifidobacterium numbers increased. Moreover, strain 06TCa19 was suggested to reach and survive in the intestines, and is, therefore, suitable for fermented dairy products and can potentially improve human defecation.


Biswas G.,University of Miyazaki | Korenaga H.,University of Miyazaki | Nagamine R.,University of Miyazaki | Kawahara S.,University of Miyazaki | And 6 more authors.
International Immunopharmacology | Year: 2013

The important role played by cytokines in host innate immunity and the interaction of subsets of immune and inflammatory cells through cytokines offer avenues for immune interventions. We investigated 16 cytokine gene responses in the Japanese pufferfish, Takifugu rubripes orally treatedwith a heat-killed lactic acid bacterium (LAB), Lactobacillus paracasei spp. paracasei (strain 06TCa22) (Lpp) isolated from a Mongolian dairy product at 1 mg g-1 bodyweight d-1 for 3 days. Additionally,we assessed superoxide anion production (SAP) and phagocytic activity (PA) of head kidney cells and resistance to Vibrio harveyi infection in treated fish. Significant up-regulation of pro-inflammatory (IL-1β, IL-6, IL-17A/F-3, TNF-aα and TNF-N), cell-mediated immunity inducing (IL-12p35, IL-12p40 and IL-18), antiviral/intra-cellular pathogen killing (I-IFN-1 and IFN-γ), anti-inflammatory (IL-10) and peripheral T cell expansion and survival controlling (IL-2, IL-7, IL-15, IL-21 and TGF-β1) cytokines was observed in the treated fish. Furthermore, significantly increased SAP, PA and pathogen resistance were observed in the treated fish compared to untreated fish. Our results indicate the enhancement of cytokine mediated immunity in T. rubripes by the use of the heat-killed Lpp as a potential immunostimulant andwould be of great use in immunomodulation trialswith the possibility tomonitor positive immune response. © 2013 Elsevier B.V. All rights reserved.


Takeshita T.,Kyushu University of Health and Welfare | Watanabe W.,Kyushu University of Health and Welfare | Toyama S.,Kyushu University of Health and Welfare | Hayashi Y.,Kyushu University of Health and Welfare | And 10 more authors.
Evidence-based Complementary and Alternative Medicine | Year: 2013

Tetrabromobisphenol A (TBBPA), a brominated flame retardant, has been found to exacerbate pneumonia in respiratory syncytial virus- (RSV-) infected mice. We examined the effect of Brazilian propolis (AF-08) on the exacerbation of RSV infection by TBBPA exposure in mice. Mice were fed a powdered diet mixed with 1% TBBPA alone, 0.02% AF-08 alone, or 1% TBBPA and 0.02% AF-08 for four weeks and then intranasally infected with RSV. TBBPA exposure increased the pulmonary virus titer and level of IFN-γ, a representative marker of pneumonia due to RSV infection, in the lungs of infected mice without toxicity. AF-08 was significantly effective in reducing the virus titers and IFN-γ level increased by TBBPA exposure. Also, AF-08 significantly reduced proinflammatory cytokine (TNF-α and IL-6) levels in the lungs of RSV-infected mice with TBBPA exposure, but Th2 cytokine (IL-4 and IL-10) levels were not evidently increased. Neither TBBPA exposure nor AF-08 treatment affected the anti-RSV antibody production in RSV-infected mice. In flow cytometry analysis, AF-08 seemed to be effective in reducing the ratio of pulmonary CD8a+ cells in RSV-infected mice with TBBPA exposure. TBBPA and AF-08 did not exhibit anti-RSV activity in vitro. Thus, AF-08 probably ameliorated pneumonia exacerbated by TBBPA exposure in RSV-infected mice by limiting excess cellular immune responses. © 2013 Tomomi Takeshita et al.


Takeda S.,Minami Nihon Rakuno Kyodo Co. | Kawahara S.,University of Miyazaki | Hidaka M.,Kyushu University of Health and Welfare | Yoshida H.,Kyushu University of Health and Welfare | And 6 more authors.
Bioscience, Biotechnology and Biochemistry | Year: 2013

We investigated 10 lactic acid bacteria strains with probiotic potential prepared from Mongolian dairy products for their ability to induce T helper type-1 (Th1) cytokine production in mouse immune cells in vitro and in vivo. Among these strains, the Lactobacillus plantarum 06CC2 strain was effective in elevating the level of interleukin (IL)-12p40 in co-culture with J774.1 cells and the levels of IL-12 and interferon (IFN)-γ in co-culture with mouse spleen cells in vitro. Oral administration of this strain augmented the gene expression of IFN-γ and IL-12p40 and enlarged the population of CD4 +, CD25+, and CD49b+ cells in the spleens of normal mice. It also significantly elevated the gene expression of IL-12 receptor β2 as well as IL-12p40 and IFN-γ in Peyer's patches. Thus oral administration of strain 06CC2 was effective in inducing Th1 cytokine production activating the Th1 immune response associated with intestinal immunity in normal mice.


Takeda S.,Minami Nihon Rakuno Kyodo Co. | Hidaka M.,Kyushu University of Health and Welfare | Yoshida H.,Kyushu University of Health and Welfare | Takeshita M.,Minami Nihon Rakuno Kyodo Co. | And 7 more authors.
Journal of Functional Foods | Year: 2014

Antiallergic activities of 10 lactic acid bacteria strains prepared from Mongolian dairy products as orally administered probiotics were examined in three murine type I allergy models (compound 48/80 stimulation, passive cutaneous anaphylaxis reaction, and ovalbumin sensitization models). Among the 10, only Lactobacillus plantarum strain 06CC2 significantly alleviated allergic symptoms in all three models and reduced the levels of total IgE, ovalbumin-specific IgE, and histamine in the sera of ovalbumin-sensitized mice. In vitro study, interferon-γ and interleukin-4 secretions from spleen cells of ovalbumin-sensitized mice administered the 06CC2 strain were significantly enhanced and suppressed, respectively, in the presence of ovalbumin. In Peyer's patches of ovalbumin-sensitized mice, strain 06CC2 significantly enhanced mRNA expressions of interferon-γ and interleukin-12 receptor β2, but suppressed that of the interleukin-4. Thus, strain 06CC2 probably promoted Th1 immunity through intestinal immunity and improved the Th1/Th2 balance in type I allergic mice, resulting in alleviation of allergic symptoms. © 2014 Elsevier Ltd.


PubMed | Minami Nihon Rakuno Kyodo Co
Type: Journal Article | Journal: Bioscience, biotechnology, and biochemistry | Year: 2013

We investigated 10 lactic acid bacteria strains with probiotic potential prepared from Mongolian dairy products for their ability to induce T helper type-1 (Th1) cytokine production in mouse immune cells in vitro and in vivo. Among these strains, the Lactobacillus plantarum 06CC2 strain was effective in elevating the level of interleukin (IL)-12p40 in co-culture with J774.1 cells and the levels of IL-12 and interferon (IFN)- in co-culture with mouse spleen cells in vitro. Oral administration of this strain augmented the gene expression of IFN- and IL-12p40 and enlarged the population of CD4(+), CD25(+), and CD49b(+) cells in the spleens of normal mice. It also significantly elevated the gene expression of IL-12 receptor 2 as well as IL-12p40 and IFN- in Peyers patches. Thus oral administration of strain 06CC2 was effective in inducing Th1 cytokine production activating the Th1 immune response associated with intestinal immunity in normal mice.


PubMed | Minami Nihon Rakuno Kyodo Co., Kyushu University of Health and Welfare, Mongolian University of Science and Technology, Mongolian Biotechnology Association and Azabu University
Type: | Journal: International immunopharmacology | Year: 2016

We previously found that Lactobacillus plantarum strain 06CC2 showed probiotic potential, and its oral administration effectively induced Th1 cytokine production and activated the Th1 immune response associated with intestinal immunity in mice. In this study, to evaluate its potential as a versatile oral adjuvant for treatment of viral infection, we assessed the immunomodulatory activity of 06CC2 on murine cutaneous herpes simplex virus type 1 (HSV-1) infection, in which a major immune defense system is a delayed-type hypersensitivity (DTH) reaction based on activation of the Th1 immune response, in relation to its oral efficacy for alleviation of herpetic symptoms. In the HSV-1 infection model, oral administration of 06CC2 (20mg/mouse) twice daily for seven days starting two days before infection was significantly effective in delaying the development of skin lesions in the early phase of infection and reducing virus yields in the brain on day 4 after infection. In addition, 06CC2 significantly augmented the DTH reaction to inactivated HSV-1 antigen and elevated interferon (IFN)- production by HSV-1 antigen from splenocytes. On day 2, natural killer (NK) cell activity was significantly elevated, and the elevation was still observed on day 4. Furthermore, gene expressions of interleukin-12 receptor 2 and IFN- in Peyers patches were augmented on day 4 by 06CC2 administration. Thus, 06CC2 was suggested to alleviate herpetic symptoms in mice in correlation with augmentation of the Th1 immune responses associated with NK cell activity through intestinal immunity. Strain 06CC2 may be a versatile oral adjuvant to activate Th1 immune response.

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